lysing matrix e tubes  (Valiant Co Ltd)

Journal: Gut Microbes

Article Title: Microbial metabolite Enterobactin impairs mitochondrial respiration and alleviates colitis

doi: 10.1080/19490976.2026.2662638

Figure Lengend Snippet: Distinct effects of bacterial siderophores (Ent, 2, 3-DHBA) and mammalian siderophore 2, 5-DHBA on mito-respiration and cell proliferation. (A) Schematic representation of the chemical structures of cyclic Ent (B) 2, 3-DHBA (C) 2, 5-DHBA (D) Seahorse XF analysis of mitochondrial respiration parameters in IEC treated with vehicle (control), Ent (25 μM), or 2, 3-DHBA (25 μM) or 2, 5-DHBA (25 μM). (E) Basal respiration, maximal respiratory capacity, and ATP production were quantified. (F) Quantification of mitochondrial ROS levels in treated cells, measured using MitoSOX TM fluorescence ( n = 5–6). (G) IEC were treated with Ent (25 µM) for 24 h to assess its impact on mRNA transcripts of tight junction proteins ( ZO-1, occludin , and claudin-1 ) involved in epithelial barrier integrity via qRT-PCR. (H) Confluent monolayers of IEC were scratched and treated with 25 µM of indicated siderophores, and wound closure was monitored and imaged at each time point using phase-contrast microscopy through Incucyte (I) Quantification of wound area showed a time-dependent inhibition of wound healing in Ent-treated cells compared to untreated controls ( n = 6–7). Data represent SEM from n = 3 biological replicates, with each biological replicate plated in 5–6 technical wells. Statistical significance was determined by one-way ANOVA with post hoc analysis (* p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001).

Article Snippet: Antibodies to OXPHOS (Catalog # 45-8099) were from Thermo Fischer, and antibodies to occludin (Catalog #40-4700) and claudin-1 (Catalog #37-4900) from Invitrogen and UCP1 (Catalog #MAB6158) from R&D Systems.

Techniques: Control, Fluorescence, Quantitative RT-PCR, Microscopy, Inhibition

Journal: Gut Microbes

Article Title: Microbial metabolite Enterobactin impairs mitochondrial respiration and alleviates colitis

doi: 10.1080/19490976.2026.2662638

Figure Lengend Snippet: Monomeric Ent, 2, 3-DHBA, but not 2, 5-DHBA, restores mucosal integrity and mitochondrial homeostasis in DSS-induced colitis. Acute colitis was induced in mice as in Figure 7. After the cessation of DSS (recovery phase) mice were given either 2, 3-DHBA or 2, 5-DHBA (500 µg/mouse per day) for 7 d and euthanized. (A) Representative H&E-stained colon sections showing epithelial damage, crypt loss, and inflammatory cells ( n = 5/group). (B) Alcian blue staining for goblet cells in the colon (C) Immunoblot analysis of tight junction proteins (occludin and claudin-1) in colonic tissue. (D) qRT-PCR analysis of genes involved in mitochondrial biogenesis peroxisome proliferator activated receptor gamma coactivator 1-alpha ( PGC-1α ) and deiodinase type 2 ( Dio2) .(E) Colonic expression of antioxidant genes (Superoxide dismutase 2 (SOD2) , glutathione peroxidase (Gpx) , and nuclear factor erythroid 2-related factor 2 (Nrf2) (F) Immunoblot showing protein involved in mitochondrial dynamics uncoupling protein 1 (Ucp1), mitofusin 1 (Mfn1), and optic atrophy type 1 (Opa1). Data is presented as mean ± SEM ( n = 5). Statistical comparisons were performed using one-way ANOVA with Tukey’s post hoc test (* p < 0.05, ** p < 0.01, and *** p < 0.001).

Article Snippet: Antibodies to OXPHOS (Catalog # 45-8099) were from Thermo Fischer, and antibodies to occludin (Catalog #40-4700) and claudin-1 (Catalog #37-4900) from Invitrogen and UCP1 (Catalog #MAB6158) from R&D Systems.

Techniques: Staining, Western Blot, Quantitative RT-PCR, Expressing